The effect mechanism of ergosterol from the nutritional mushroom Leucocalocybe mongolica in breast cancer cells: Protein expression modulation and metabolomic profiling using UHPLC-ESI-Q.

The ergosterol from mushrooms has gained significant ethnopharmacological importance in various cultures, including China, Japan, and Europe. This compound has been found to possess immune-boosting and anti-inflammatory properties, making it useful in the treatment of immune disorders. In this study, we focused on investigating the potential anticancer properties of ergosterol isolated from the edible mushroom Leucocalocybe mongolica in breast cancer cell lines. The ergosterol was purified and identified using advanced analytical techniques such as ESI-MS and NMR. We conducted cell proliferation assays on 4 T1 breast cancer cells to assess the cytotoxic effects of ergosterol. Furthermore, we analyzed the transcription levels of BAX, caspase-7, BCL-2, STAT-3, and PARP proteins using real-time PCR and Western blot analysis. Additionally, we employed non-targeted ultra-high-performance liquid chromatography and high-resolution mass spectrometry (UPLC-MS/MS) to study the potential mechanisms underlying the anticancer effects of ergosterol at the metabolomics level. The results demonstrated a significant reduction in cell viability and the induction of apoptosis upon treatment with ergosterol, especially at higher concentrations (P < 0.05). Moreover, ergosterol affected the expression of cancer-related genes, upregulating pro-apoptotic proteins such as BAX, caspase-7, and PARP, while downregulating the anti-apoptotic proteins BCL-2 and STAT-3 (P < 0.05). Western blot analysis confirmed these findings and provided further evidence of ergosterol's role in inducing apoptosis. Metabolomics analysis revealed substantial changes in pathways related to amino acid, antioxidant, and carbohydrate metabolism. In conclusion, our study demonstrates that ergosterol exhibits anticancer effects by inducing apoptosis and modulating metabolic pathways in breast cancer cells.

Merremia emarginata Extract Potentiates the Inhibition of Human Colon Cancer Cells (HT-29) via the Modulation of Caspase-3/Bcl-2-Mediated Pathways.

Background This study investigates Merremia emarginata's curative effectiveness against colon cancer cells. M. emarginata, often known as Elika jemudu, is a Convolvulaceae family plant. The inhibitory ability of anticancer herbal extracts against cancer cell growth and mediators is tested.  Aim This study aims to evaluate the potent anticancer activity of M. emarginata against colon cancer cell line (HT-29). Materials and methods M. emarginata leaves were gathered and processed using solvent extraction. Anticancer activity on colon cancer cells was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test and cysteine aspartic acid protease-3 (caspase 3), B-cell lymphoma 2 (Bcl-2), and B-cell lymphoma-extra large (Bcl-xL) mRNA expressions. The data was reported as the mean ± SD of three separate experiments done in triplicate. The statistical analysis was carried out using one-way analysis of variance (ANOVA), with a p-value less than 0.05 indicating statistical significance. Results The cell viability test showed a gradual decrease in cell growth and proliferation as the concentration increased. The ethanolic extract of M. emarginata was found to be cytotoxic against colon caller cell lines. The extract was able to induce apoptosis of cancer as revealed by Bcl-2, Bcl-xL, and caspase-3 (p<0.05 and p<0.001) signaling pathways. Conclusion M. emarginata extracts showed good anticancer activity against colon cancer cell lines. Further work is required to establish and identify the chemical constituent responsible for its anticancer activity.

Cell viability and cytotoxicity assays: Biochemical elements and cellular compartments.

Cell viability and cytotoxicity assays play a crucial role in drug screening and evaluating the cytotoxic effects of various chemicals. The quantification of cell viability and proliferation serves as the cornerstone for numerous in vitro assays that assess cellular responses to external factors. In the last decade, several studies have developed guidelines for defining and interpreting cell viability and cytotoxicity based on morphological, biochemical, and functional perspectives. As this domain continues to experience ongoing growth, revealing new mechanisms orchestrating diverse cell cytotoxicity pathways, we suggest a revised classification for multiple assays employed in evaluating cell viability and cell death. This classification is rooted in the cellular compartment and/or biochemical element involved, with a specific focus on mechanistic and essential aspects of the process. The assays are founded on diverse cell functions, encompassing metabolic activity, enzyme activity, cell membrane permeability and integrity, adenosine 5'-triphosphate content, cell adherence, reduction equivalents, dye inclusion or exclusion, constitutive protease activity, colony formation, DNA fragmentation and nuclear splitting. These assays present straightforward, reliable, sensitive, reproducible, cost-effective, and high-throughput approaches for appraising the effects of newly formulated chemotherapeutic biomolecules on the cell survival during the drug development process.

Membranes composed of poly(lactic acid)/poly(ethylene glycol) and Ora-pro-nóbis (Pereskia aculeata Miller) extract for dressing applications.

Wounds are considered one of the most critical medical conditions that must be managed appropriately due to the psychological and physical stress they cause for patients, as well as creating a substantial financial burden on patients and global healthcare systems. Nowadays, there is a growing interest in developing nanofiber mats loaded with varying plant extracts to meet the urgent need for advanced wound ressings. This study investigated the development and characterization of poly(lactic acid) (PLA)/ poly(ethylene glycol) (PEG) nanofiber membranes incorporated with Ora-pro-nóbis (OPN; 12.5, 25, and 50 % w/w) by the solution-blow-spinning (SBS) technique. The PLA/PEG and PLA/PEG/OPN nanofiber membranes were characterized by scanning electron microscopy (SEM), thermal properties (TGA and DSC), Fourier transform infrared spectroscopy (FTIR), contact angle measurements and water vapor permeability (WVTR). In addition, the mats were analyzed for swelling properties in vitro cell viability, and fibroblast adhesion (L-929) tests. SEM images showed that smooth and continuous PLA/PEG and PLA/PEG/OPN nanofibers were obtained with a diameter distribution ranging from 171 to 1533 nm. The PLA/PEG and PLA/PEG/OPN nanofiber membranes showed moderate hydrophobicity (~109-120°), possibly preventing secondary injuries during dressing removal. Besides that, PLA/PEG/OPN nanofibers exhibited adequate WVTR, meeting wound healing requirements. Notably, the presence of OPN gave the PLA/PEG membranes better mechanical properties, increasing their tensile strength (TS) from 3.4 MPa (PLA/PEG) to 5.3 MPa (PLA/PEG/OPN), as well as excellent antioxidant properties (Antioxidant activity with approximately 45 % oxidation inhibition). Therefore, the nanofiber mats based on PLA/PEG, especially those incorporated with OPN, are promising options for use as antioxidant dressings to aid skin healing.

Melatonin improves the viability and ultrastructure of bovine oocyte-granulosa complexes of in vitro cultured early antral follicles.

This study aims to investigate the effects of melatonin on follicular growth, viability and ultrastructure, as well as on the levels of mRNA for antioxidant enzymes, reactive oxygen species (ROS) and meiotic progression in oocytes from in vitro cultured bovine early antral follicles. To this end, isolated early antral follicles (500-600 µm) were cultured in TCM-199+ alone or supplemented with 10-6 , 10-7 or 10-8 M melatonin at 38.5°C with 5% CO2 for 8 days. Follicle diameters were evaluated at days 0, 4 and 8 of culture. At the end of culture, ultrastructure, chromatin configuration, viability (calcein-AM and ethidium homodimer-1 staining), and the levels of ROS and mRNA for catalase (CAT), superoxide dismutase (SOD) and peroxiredoxin 6 (PRDX6) and glutathione peroxidase (GPx) were investigated in oocyte-granulosa cell complexes (OGCs). The results showed that early antral follicles cultured with 10-6 and 10-8 M melatonin had a progressive and significant increase in their diameters throughout the culture period (p < .05). Additionally, oocytes from follicles cultured with 10-7 or 10-8 M melatonin had increased fluorescence for calcein-AM, while those cultured with 10-6 or 10-7 M had reduced fluorescence for ethidium homodimer-1. Different from follicles cultured in other treatments, those cultured with 10-8 M melatonin had well-preserved ultrastructure of oocyte and granulosa cells. Melatonin, however, did not influence the levels of ROS, the mitochondrial activity, oocyte meiotic resumption and expression mRNA for SOD, CAT, GPX1 and PRDX6. In conclusion, the presence of 10-8 M melatonin in culture medium improves viability and preserves the ultrastructure of oocyte and granulosa cells of early antral follicles cultured in vitro.

Optimization of Antioxidant Activity of Compounds Generated during Ginseng Extract Fermentation Supplemented with Lactobacillus.

Ginseng holds high medicinal and cosmetic value, with stem and leaf extracts garnering attention for their abundant bioactive ingredients. Meanwhile, fermentation can enhance the effectiveness of cosmetics. The aim of this study was to optimize ginseng fermentation to produce functional cosmetics. Ginseng stem and leaf extracts were fermented with five different strains of lactic acid bacteria. Using 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl radical (·OH), and superoxide anion (O2·-) scavenging activities as indicators, the fermentation process was optimized via response surface methodology. Finally, validation of the antioxidant activity of the optimized fermentation broth was performed using human skin cells (HaCaT and BJ cells). Based on the antioxidant potency composite comprehensive index, Lactiplantibacillus plantarum 1.140 was selected, and the optimized parameters were a fermentation time of 35.50 h, an inoculum size of 2.45%, and a temperature of 28.20 °C. Optimized fermentation boosted antioxidant activity: DPPH scavenging activity increased by 25.00%, ·OH by 94.00%, and O2·- by 73.00%. Only the rare ginsenoside Rg5 showed a substantial rise in content among the 11 ginsenosides examined after fermentation. Furthermore, the flavonoid content and ·OH scavenging activity were significantly negatively correlated (r = -1.00, p < 0.05), while the Rh1 content and O2·- scavenging activity were significantly positively correlated (r = 0.998, p < 0.05). Both the 0.06% (v/v) and 0.25% (v/v) concentrations of the optimized broth significantly promoted cell proliferation, and notable protective effects against oxidative damage were observed in HaCaT cells when the broth was at 0.06%. Collectively, we demonstrated that ginseng fermentation extract effectively eliminates free radicals, preventing and repairing cellular oxidative damage. This study has identified new options for the use of fermented ginseng in functional cosmetics.

Chemical Analysis and Biological Activities of Extracts Isolated from Symbiotic L. japonicus Plants.

Plants produce a wide variety of secondary metabolites, including compounds with biological activities that could be used for the treatment of human diseases. In the present study, we examined the putative production of bioactive molecules in the legume plant Lotus japonicus, which engages into symbiotic relationships with beneficial soil microorganisms. To monitor the production of secondary metabolites when the plant develops beneficial symbiotic relationships, we performed single and double inoculations with arbuscular mycorrhizal fungi (AMF) and nitrogen-fixing Rhizobium bacteria. Plant extracts from non-inoculated and inoculated plants were chemically characterized and tested for anti-proliferative, apoptotic, and anti-inflammatory effects on human HEK-293 cells. Both shoot and root extracts from non-inoculated and inoculated plants significantly reduced the HEK-293 cell viability; however, a stronger effect was observed when the root extracts were tested. Shoot and root extracts from Rhizobium-inoculated plants and shoot extracts from AMF-inoculated plants showed apoptotic effects on human cells. Moreover, both shoot and root extracts from AMF-inoculated plants significantly reduced TNFα-induced NF-κB transcriptional activity, denoting anti-inflammatory activity. These results suggest that symbiotic L. japonicus plants are enriched with metabolites that have interesting biological activities and could be further explored for putative future use in the pharmaceutical sector.

The potential of Rhodobacter sphaeroides extract as an alternative supplement for cell culture systems.

It is essential to identify suitable supplements that enhance cell growth, viability, and functional development in cell culture systems. The use of fetal bovine serum (FBS) has been common, but it has limitations, such as batch-to-batch variability, ethical concerns, and risks of environmental contamination. In this study, we explore the potential of Rhodobacter sphaeroides extract, derived from a probiotic photosynthetic bacterium, as an alternative supplement. Our results demonstrate that the extract from R. sphaeroides significantly improves various aspects of cell behavior compared to serum-free conditions. It enhances cell growth and viability to a greater extent than FBS supplementation. Additionally, the extract alleviates oxidative stress by reducing intracellular levels of reactive oxygen species and stimulates lysosomal activity, contributing to cellular processes. The presence of abundant amino acids, glycine and arginine, in the extract may play a role in promoting cell growth. These findings emphasize the potential of R. sphaeroides extract as a valuable supplement for cell culture, offering advantages over the use of FBS.IMPORTANCEThe choice of supplements for cell culture is crucial in biomedical research, but the widely used fetal bovine serum (FBS) has limitations in terms of variability, ethics, and environmental risks. This study explores the potential of an extract from Rhodobacter sphaeroides, a probiotic bacterium, as an alternative supplement. The findings reveal that the R. sphaeroides extract surpasses FBS in enhancing cell growth, viability, and functionality. It also mitigates oxidative stress and stimulates lysosomal activity, critical for cellular health. The extract's abundance of glycine and arginine, amino acids with known growth-promoting effects, further highlights its potential. By providing a viable substitute for FBS, the R. sphaeroides extract addresses the need for consistent, ethical, and environmentally friendly cell culture supplements. This research paves the way for sustainable and reliable cell culture systems, revolutionizing biomedical research and applications in drug development and regenerative medicine.

Polyfunctional sugar-free white chocolate fortified with Lacticaseibacillus rhamnosus GG co-encapsulated with beet residue extract (Beta vulgaris L.).

Chocolate is a worldwide consumed food. This study investigated the fortification of sugar-free white chocolate with Lacticaseibacillus rhamnosus GG microcapsule co-encapsulated with beet residue extract. The chocolates were evaluated for moisture, water activity, texture, color properties, melting, physicochemical, and probiotic stability during storage. Furthermore, the survival of L. rhamnosus GG and the bioaccessibility of phenolic compounds were investigated under in vitro simulated gastrointestinal conditions. Regarding the characterization of probiotic microcapsules, the encapsulation efficiency of L. rhamnosus GG was > 89 % while the encapsulation efficiency of phenolic compounds was > 62 %. Chocolates containing probiotic microcapsules were less hard and resistant to breakage. All chocolates had a similar melting behavior (endothermic peaks between 32.80 and 34.40 °C). After 120 days of storage at 4 °C, probiotic populations > 6.77 log CFU/g were detected in chocolate samples. This result demonstrates the potential of this matrix to carry L. rhamnosus GG cells. Regarding the resistance of probiotic strains during gastric simulation, the co-encapsulation of L. rhamnosus GG with beet extract contributed to high counts during gastrointestinal transit, reaching the colon (48 h) with viable cell counts equal to 11.80 log CFU/g. Finally, one of our main findings was that probiotics used phenolic compounds as a substrate source, which may be an observed prebiotic effect.

In-vitro Antioxidant Activity and Flow Cytometric Analysis of Simarouba glauca DC Bark Extract Induced Apoptosis in Triple Negative Breast Cancer Cells.

OBJECTIVE: Ethnomedicinally Simarouba glauca DC is an important plant containing major class of phenols and terpenoids as bioactive compounds. The present study focuses on the evaluation of the anticancer effects of S. glauca bark UAE-EA (Ultrasonicator Assisted Extraction - Ethyl Acetate) fraction (SG-Fraction) against MDA-MB-231 triple negative breast cancer cell lines. METHODS: UAE-EA technique was used for the extraction of phytochemicals from S. glauca bark. Fractionation method was carried out to obtain Ethyl acetate fraction and PPS, TPC, and DPPH assays were performed to characterize the extract. MTT assay was then applied to analyse the viability of cells and MMP assay to confirm the initiation of drug induced apoptosis. Apoptotic morphology and quantification were assessed by DAPI and Annexin V/propidium iodide (PI) staining. RESULTS: UAE yielded 53g of crude extract in methanol. 16g Ethyl acetate fraction was obtained from fractionation. Phytoconstituents such as alkaloids, phenols, flavonoids, and triterpenoids were detected. The TPC was 278.65 mg GAE/100ml. The SG-Fraction showed maximum 66.38% RSA at 200 µg/ml and IC50 value was 101.72 µg/ml. MMP confirmed the induction of apoptosis. DAPI showed the reduction of nuclei with bright chromatin condensation, blebbing, nuclear fragmentation and apoptotic bodies. Annexin-V FITC/PI study showed 59.48% apoptosis induction. This fraction showed a similar trend of antioxidant effect as compared to ascorbic acid but, prominently lower cell viability than Camptothecin (P<0.005). In line with higher TPC in the SG-fraction, free radical scavenging activity was increased (r = 0.098**, p=0.002) and cell viability was reduced significantly (r = -0.097*** p<0.01). CONCLUSION: These results indicate that UAE-EA fraction of S. glauca bark inhibits the growth of MDA-MB-231 cells and can be considered for further neo-adjuvant chemotherapy drug research.

Porous scaffold hydroxyapatite from sand lobster shells (Panulirus homarus) using polyethylene oxide/chitosan as polymeric porogen for bone tissue engineering.

The hydroxyapatite (HAp; Ca10 (PO4 )6 (OH)2 )) has good biocompatibility, bioactivity, and osteoconductivity as a bone implant because the main inorganic mineral of human bone is HAp. The use of scaffold HAp from biogenic resources that contain high calcium and polymer as a pore forming agent to support bone growth is a longstanding area of interest. In this study, porous scaffolds based on HAp were synthesized from sand lobster (SL; Panulirus homarus) shells as a source of calcium using the porogen leaching method with polyethylene oxide (PEO) and chitosan (Chs) as polymeric porogen. The present study aims to synthesize HAp derived from SL shells and evaluate the effect variations of PEO on the physicochemical properties of the scaffold and cytotoxicity in cell viability assay. Briefly, the SL shell powder was calcinated with temperature variations of 600°C, 800°C, and 1000°C for 6 h. Based on the characterization, it was shown that 1000°C was the optimum calcination temperature for SL shells to synthesize HAp using the precipitation method. The characterization results of HAp using energy dispersive x-ray (EDX) revealed that the molar ratio of Ca/P was 1.67. The Fourier transform infrared (FTIR) and x-ray diffractometer (XRD) spectral patterns indicated that HAp had been successfully synthesized with minor ß-tricalcium phosphate (ß-TCP), a calcium phosphate with high biocompatibility. Porous scaffolds were synthesized by varying the concentration of PEO at 0, 5, 10, and 15 wt %. Physicochemical analysis revealed that a higher concentration of PEO affected decreased crystallinity and compressive strength, but on the other hand, the porosity and pore sizes increased. Based on the physicochemical analysis, the synthesized porous scaffold showed that HAp/PEO/Chs 15 wt % had the most potential as a scaffold for biomedical applications. MTT Assay, after 24 h incubation, revealed that the scaffold was safe for use at low concentrations on the MC3T3E1 osteoblast cells, with a percentage of cell viability of 83.23 ± 3.18% at 23.4375 µg/mL. Although the cell viability decreased at higher concentrations, the HAp/PEO/Chs 15 wt % scaffold was cytocompatible with the cells. Thus, in the present study, HAp/PEO/Chs 15 wt % was the best scaffold based on pore structure, chemical composition, mechanical and crystalographic properties and cell viability.

Synthesis of homogeneous spherical selenium nanoparticles through a chemical method for cancer therapy applications.

Oxidative stress is associated with pathologies affecting various organs or metabolic pathways. Thus, targeting oxidative stress might represent a valid therapeutic option. Selenium nanoparticles (SeNPs) are reported to exert antioxidant effects by many mechanisms. Our purpose was to assess in vitro on normal (MRC-5) and cancer (PANC-1) cell lines the potential of SeNPs for inducing cytotoxicity and redox modulation. They were synthesized through a chemogenic method and characterized through advanced microscopy techniques. SeNPs were spherical, with 100 nm average diameters and low dimension variability. Cancer and normal cells were exposed for 24 h to different concentrations of SeNPs ranging from 1 to 25 µg/mL. According to the LDH and MTT assay results, SeNPs treatment caused a more pronounced decrease in cancer cell viability compared to normal cells, suggesting a possible therapeutic benefit on tumors, thus supporting the hypothesis of therapeutic use of SeNPs with the benefit of cell type selectivity. Neither an elevation nor an inhibition of intracellular ROS production was detected in MRC-5 cells exposed to concentrations between 1 and 25 µg/mL SeNPs. The results of this study suggest that SeNPs could represent potential candidate for treatment of cancer, especially pancreatic adenocarcinoma.

Green Synthesis of Chitosan-Capped Gold Nanoparticles Using Salvia officinalis Extract: Biochemical Characterization and Antimicrobial and Cytotoxic Activities.

Increasing antimicrobial resistance to the action of existing antibiotics has prompted researchers to identify new natural molecules with antimicrobial potential. In this study, a green system was developed for biosynthesizing gold nanoparticles (BAuNPs) using sage (Salvia officinalis L.) leaf extract bioconjugated with non-toxic, eco-friendly, and biodegradable chitosan, forming chitosan/gold bioconjugates (Chi/BAuNPs). Characterization of the BAuNPs and Chi/BAuNPs conjugates takes place using transmission electron microscopy (TEM), X-ray spectra, Fourier transform infrared (FT-IR) spectroscopy, and zeta potential (Z-potential). The chemical composition of S. officinalis extract was evaluated via gas chromatography/mass spectrometry (GC/MS). This study evaluated the antioxidant and antimicrobial activities of human pathogenic multidrug-resistant (MDR) and multisensitive (MS) bacterial isolates using the agar diffusion method. Chi/BAuNPs showed inhibition of the MDR strains more effectively than BAuNPs alone as compared with a positive standard antibiotic. The cytotoxicity assay revealed that the human breast adenocarcinoma cancer cells (MCF7) were more sensitive toward the toxicity of 5-Fu + BAuNPs and 5-Fu + Chi/BAuNPs composites compared to non-malignant human fibroblast cells (HFs). The study shows that BAuNPs and Chi/BAuNPs, combined with 5-FU NPs, can effectively treat cancer at concentrations where the free chemical drug (5-Fu) is ineffective, with a noted reduction in the required dosage for noticeable antitumor action.

Steroidal Alkaloids from Food Waste of Tomato Processing Inhibit Neuroblastoma Cell Viability.

Nowadays, there is considerable attention toward the use of food waste from food processing as possible sources of compounds with health properties, such as anticancer activity. An example is tomato processing, which is responsible for generating a remarkable amount of waste (leaves, peel, seeds). Therefore, our goal was to evaluate the potential anticancer property of tomato extracts, in particular "Datterino" tomato (DT) and "Piccadilly" tomato (PT), and to study their phytochemical composition. Liquid chromatography with tandem mass spectrometry (LC/MS-MS) results showed that these extracts are rich in alkaloids, flavonoids, fatty acids, lipids, and terpenes. Furthermore, their potential anticancer activity was evaluated in vitro by MTT assay. In particular, the percentage of cell viability was assessed in olfactory ensheathing cells (OECs), a particular glial cell type of the olfactory system, and in SH-SY5Y, a neuroblastoma cell line. All extracts (aqueous and ethanolic) did not lead to any significant change in the percentage of cell viability on OECs when compared with the control. Instead, in SH-SY5Y we observed a significant decrease in the percentage of cell viability, confirming their potential anticancer activity; this was more evident for the ethanolic extracts. In conclusion, tomato leaves extracts could be regarded as a valuable source of bioactive compounds, suitable for various applications in the food, nutraceutical, and pharmaceutical fields.

Efficacy of Bee Products (Anzer Honey, Pollen and Propolis) in Detection and Healing of Damage Induced by Antidiabetic Drug Vildagliptin/Metformin Hydrochloride in Healthy Human Pancreatic Cells: Cytotoxic, Genotoxic and Biochemical Studies.

BACKGROUND AND OBJECTIVE: Although drugs are powerful therapeutic agents, they have a range of side effects. These side effects are sometimes cellular and not clinically noticeable. Vildagliptin/metformin hydrochloride is one of the most widely used oral antidiabetic drugs with two active ingredients. In this study, we investigated its harmful effects on the metabolic activation system in healthy human pancreatic cells "hTERT-HPNE", and we aimed to improve these harmful effects by natural products. To benefit from the healing effect, we used the unique natural products produced by the bees of the Anzer Plateau in the Eastern Black Sea Region of Turkey. METHODS: Cytotoxic and genotoxic effects of the drug were investigated by different tests, such as MTT, flow cytometry-apoptosis and comet assays. Anzer honey, pollen and propolis were analyzed by gas chromatography/mass spectrometry (G/C-MS). A total of 19 compounds were detected, constituting 99.9% of the samples. RESULTS: The decrease in cell viability at all drug concentrations was statistically significant compared to the negative control (P<0.05). A statistically significant decrease was detected in the apoptosis caused by vildagliptin/metformin hydrochloride with the supplementation of Anzer honey, pollen and propolis in hTERT-HPNE cells (P<0.05). CONCLUSION: This study can contribute to other studies testing the healing properties of natural products against the side effects of oral antidiabetics in human cells. In particular, Anzer honey, pollen and propolis can be used as additional foods to maintain cell viability and improve heal damage and can be evaluated against side effects in other drug studies.

The effect of environmental stressors on the anticancer potential of Amaranthus hypochondriacus aqueous extracts and fractions.

Some Amaranthus species have been shown to have pharmacological properties such as activity against cancer, and it is also used as a traditional herbal medicine in many rural parts of the world. The (3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide assay was used as a screening tool to determine the approximate cell viability inhibitory concentrations of methanol and aqueous crude extracts of Amaranthus spp. The extracts were screened using small-cell lung cancer (H69V), hepatocellular carcinoma (HepG2/C3A) and non-cancerous kidney cells (Vero) cell lines. Viability was assessed following exposure to a series of concentrations of each extract and A. hypochondriacus showed cytotoxicity of 70.55 µg/mL against H69V with a Si index of 1.8. The fractionated aqueous extract of 40 °C-treated A. hypochondriacus under well-watered conditions had a higher viability inhibition on H69V and Vero cell lines compared to the A. caudatus, A. cruentus and A. spinosus crude extracts. In conclusion, A. hypochondriacus could serve as a potential source of anticancer phytoconstituents for drug development.

Viability of human dental pulp stem cells: The potential of L-arginine-based culture media.

Dental pulp is built by proteins that have various roles in the biological process of pulp, such as structural protein, regulation protein, and catalytic protein. L-arginine, an amino acid and one of the building blocks of proteins, regulates pro-inflammatory and anti-inflammatory activity. Therefore, L-arginine-based culture has potential to promote dental pulp regeneration. This study aimed to investigate the potential of L-arginine-based culture in improving the viability of human dental pulp stem cells (hDPSCs). We evaluated the viability of hDPSCs in culture media supplemented with different concentrations of L-arginine amino acid (250, 300, 350, and 400 µmol/L) and Dulbecco's Modified Eagle Medium plus fetal bovine serum 10% (control) using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay after 24-h incubation time. Statistical analysis was conducted using a one-way analysis of variance and post hoc least significant difference test. In qualitative analysis, the 4´, 6-diamidino-2-phenylindole staining method was used. The evaluation has shown a significant result when 250, 300, and 350 µmol/L concentration of L-arginine amino acid culture media compared with control, and 400 µmol/L has the best result and was not significantly different with control toward viability of hDPSCs.

Effect of the Pulsed Electric Field Treatment on Physical, Chemical and Structural Changes of Vacuum Impregnated Apple Tissue in Aloe Vera Juices.

Vacuum impregnation (VI) stands as a diffusion-driven food processing method that has found recent application within the food industry, particularly for the cold formulation of fortified food products. Pulsed electric field (PEF) treatment can affect the food structure, influencing therefore the mass transfer phenomena during the further processing. Thus, the study aimed at investigating the effect of PEF treatment on selected physicochemical properties of vacuum-impregnated apples. Apple slices were vacuum impregnated with aloe vera juice solution with or PEF treatment at different intensities (125, 212.5 or 300 V/cm). The PEF was applied as a pretreatment-applied before the VI process as well as posttreatment-applied after the VI process. The VI process with aloe vera juice resulted in a sample weight increase of over 24% as well as structural changes, partial cell viability loss and color alteration. In addition, the decrease of bioactive compounds was observed, while antioxidant activity remained at a similar level as in raw material. PEF treatment adversely affected vacuum impregnation efficiency, causing microstructural changes and cell viability loss. Additionally, chemical composition modifications were evident through thermogravimetric analysis (TGA) and Fourier Infrared Spectroscopy (FTIR) analyses. Tissue hardness decreased significantly due to structural damage and caused high leakage from plant tissue, which resulted in hindering saturation with aloe vera juice during the VI process. Additionally, reduced bioactive substance content after PEF treatment was observed and the VI process did not restore apple samples of the bioactive compounds from aloe vera juice.

Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts.

As byproducts of essential oil distillation, hydrolates are used in natural cosmetics/biomedicine due to their beneficial skin effects. However, data on their safety with relevant biological targets, such as human skin cells, are scarce. Therefore, we have tested nine hydrolates from the Lamiaceae family with skin fibroblasts that are responsible for extracellular collagenous matrix builds. Thyme, oregano, and winter savoury hydrolates showed several times higher total phenolics, which correlated strongly with their radical scavenging and antioxidative capacity; there was no correlation between their viability profiles and the reducing sugar levels. No proteins/peptides were detected. All hydrolates appeared safe for prolonged skin exposure except for 10-fold diluted lavender, which showed cytotoxicity (~20%), as well as rosemary and lavandin (~10%) using viability, DNA synthesis, and cell count testing. Clary sage, oregano, lemon balm, and thyme hydrolates (10-fold diluted) increased fibroblast viability and/or proliferation by 10-30% compared with the control, while their viability remained unaffected by Mentha and winter savoury. In line with the STITCH database, increased viability could be attributed to thymol presence in oregano and thyme hydrolates in lemon balm, which is most likely attributable to neral and geranial. The proliferative effect of clary sage could be supported by alpha-terpineol, not linalool. The major volatile organic compounds (VOCs) associated with cytotoxic effects on fibroblasts were borneol, 1,8-cineole, and terpinene-4-ol. Further research with pure compounds is warranted to confirm the roles of VOCs in the observed effects that are relevant to cosmetic and wound healing aspects.